Interference-Fringe Microscope for Thick-Sample Super-Resolution
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Solution Overview
Problem
Existing scanning type microscopes face challenges in achieving high signal-to-noise ratio (SNR) and super-resolution imaging in thick fluorescent samples, particularly in enhancing contrast and resolving power.
Innovation Solution
A microscope design that utilizes a scanning type fluorescence microscope with an illumination optical system generating interference fringes, combined with a detection system and image processor to enhance contrast and resolution by adjusting polarization and scanning directions, and an image processing method to reconstruct super-resolved images.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional widefield fluorescence microscopy is used, then the imaging process is simple, but the contrast and resolution are insufficient in thick samples
Solution Approach 1:
The patent employs periodic scanning of line-shaped illumination across the sample in multiple directions. The illumination beam scans back and forth in the first direction, then scans in the second direction perpendicular to the first, creating periodic illumination patterns that enable super-resolution imaging through structured illumination microscopy principles
Solution Approach 2:
The patent divides the illumination into discrete line segments that are scanned across the sample. The line-shaped illumination is segmented into multiple scan positions, and the detection device captures fluorescence from each segmented region sequentially, building up the complete image through integration of these segmented measurements
2Measurement precision
If line-shaped illumination scanning is used, then super-resolution is achieved, but the signal-to-noise ratio decreases in thick samples
Solution Approach 1:
The patent merges the signals from multiple scanning directions by combining the first image data obtained from scanning in the first direction with the second image data obtained from scanning in the second direction. This merging of multi-directional data increases the signal-to-noise ratio while preserving the super-resolution capability
Solution Approach 2:
The patent introduces a second scanning dimension perpendicular to the first scanning direction. By scanning in both the first direction and the second direction, the system collects image data from multiple dimensional perspectives, which when combined, improves the signal-to-noise ratio while maintaining super-resolution
3Measurement precision
If multiple scanning directions are used, then contrast is enhanced, but the observation time increases
Solution Approach 1:
The patent uses periodic scanning in multiple directions to enhance contrast through structured illumination. The regular back-and-forth scanning patterns in both first and second directions create consistent illumination modulations that improve contrast while maintaining efficient time utilization
Solution Approach 2:
The patent maintains continuous useful action by seamlessly transitioning between scanning in the first direction and scanning in the second direction. The scanning process continues without interruption, alternating between directions to collect complementary image data that enhances contrast
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution enhances the contrast and resolution of fluorescence microscopy, enabling high SNR and super-resolution imaging in thick samples by optimizing the interference fringes and polarization states, improving the image reconstruction process.
Implementation Method 1
an illumination optical system that scans the sample with interference fringes generated by interfering two or more light fluxes
Implementation Method 2
a fluorescent substance in the sample is excited by the interference fringes of excitation light, and fluorescence from the sample is detected
Data Source
Figure 1
Figure 2(A)~2(C)
Figure 3(A)~3(C)
AI summary
To acquire an image of a sample. A microscope includes: an illumination optical system that includes a light flux splitter that splits light from a light source into a plurality of light fluxes, and scans a sample in a plurality of directions with interference fringes generated by interference of at least part of the light fluxes split by the light flux splitter; a detection optical system on which light from the sample is incident; a detection device that includes a plurality of detectors that detect the light from the sample via the detection optical system; and an image processor that generates an image using detection results of two or more of the detectors of the detection device.